6-fluoro steroids and process



United States Patent Ofiice 2,838,497

Patented June 10, 1958 corticoid activity and 2.4 times the anti-inflammatory 2 838 497 activity of hydrocortisone, while the Zl-acetate is eleven times more active than hydrocortisone as a glucocorticoid 6-FLUOR0 STEROIDS AND PROCESS and 8.7 times more active as an anti-inflammatory agent.

5 Both compounds have a favorable effect on body electrolyte balance. In addition, these compounds are useful as intermediates for the production of l-dehydro ana- George B. Spero, Kalamazoo, and John A. Hogg, Kalamazoo Township, Kalamazoo County, Mich., assignors to The Upjohn Company, Kalamazoo, Mich., a corpomfion of Michigan logues which are of particular importance because they possess marked anti-rheumatoid arthritic, anti-inflamma- No Drawing lfP November 1957 lo tory and glucocorticoid activities. For example, l-dehy- Sena 699,453 dro-6a-fluorohydrocortisone has been found to exhibit 36 Claims. (C| 26o 239 55) approximately eighty times the glucocorticoid activity and five to twenty times the anti-inflammatory activity of hydrocortisone, while the Zl-acetate is 100 times more This invention relates to 6-fluorohydrocortisone and 15 active than hydrocortisone as a glucocorticoid. The com- 2l-esters thereof, to 6-fiuorocortisone and 21-esters therepounds are useful in the treatment of inflammatory conof, to novel intermediates in the production thereof and ditions of the skin, eyes and ears of humans and valuable to processes for the production of the novel compounds domestic animals, contact dermatitis and other allergenic and the novel intermediates. reactions. The compounds can be administered in con- This application is a continuation-in-part of applica- 20 ventional dosage forms such as pills, tablets and capsules tion Serial No. 519,632, filed July 1, 1955, and of apfor oral use or in conventional liquid forms as are used plication Serial No. 634,828, filed January 18, 1957. with natural and synthetic corical steroid hormones for It has been discovered that 6-fiuorohydrocortisone and injection use. For topical use they can be administered the 21esters thereof and fi-fiuorocortisone and ZI-esters in the form of ointments, creams, lotions and the like thereof possess valuable anti-rheumatoid arthritic, antiwith or without coacting antibiotics, germicides and the inflammatory and glucocorticoid activity to a marked like. degree. Thus, for example, 6a-fluorohydrocortisone has The compounds of this invention can be prepared in been found to exhibit approximately nine times the glucoaccordance with the following scheme of reactions:

OH: OH:

OH; CH;

RI! RI! H-0 lH-O ham-o r tomb-0 i 11 OH OH ?Hz01kfi OH E0 no OH; on;

RI! RI! l (IE-0 311-0 H ..-0 f v (on n-o l 0'11 2) 0H onion'" l---oH no no CH3 CH8 m o v wherein R is hydroxy or keto, R and R" are hydrogen or loWer-alkyl, n is the whole number one or two, Ac is the acyl radical of an organic carboxylic acid, preferably a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive, and R is hydrogen or Ac as defined above. The term lower-alky], when used herein, refers to an alkyl radical of from one to eight carbon atoms, inclusive, e. g., methyl, ethyl, propyl, hutyl. amyl, hexyl, heptyl, ethylhexyl, and octyl.

The above reactions are described and exemplified hereinbelow. It will be understood by those skilled in the art. nevertheless, that the specific order of steps may be inverted or transposed or otherwise varied to suit the purposes of economics, convenience, or the like.

The preferred compounds containing the l7(20)-double bond have the cis configuration, because the cis isomers can ultimately be converted in higher yields in the oxidative hydroxylation step than is ordinarily possible with the trans isomer. it should be understood, however, that the trans isomer or mixtures of the cis and trans isomers can be used with satisfactory results.

in carrying out the epoxidation step shown above, a S-itetal of 3-keto-1loxygenated-4,l7(20)-pregnadiene-2lcarbonyloxy steroid (l), which can be prepared as disclosed in U. S. Patent 2,707,l84. preferably the 3-ethylene ketal of methyl 3,11-diketo-4,17(20)-[cis]--pregnadien-2loate. is epoxidized with a peracid, e. g., peracetic or perhenzoic, or other known epoxidizing agents, to produce the corresponding 5:6-oxide (ll). A mixture of both the aand doxides is produced in this epoxidation reaction, and the mixture can be separated by chromatographic or crystallization techniques known in the art.

OHnOAc VII In the oxide opening step, a B-ketalized 3-keto-5a,6txoxido-l1-oxygenated-l7(20)-pregnene 21 carbonyloxy steroid (ii), is reacted with hydrogen fluoride to open the oxide ring and produce the corresponding 3-ketalized 3keto-5hydroxy-6-luoro-l l-oxygenated-l7(20) pregnene-Zl-carbonyloxy steroid (lll). This epoxide opening step is ordinarily carried out at temperatures between about minus forty and plus fifty degrees centigrade, the preferred limits being between about zero and 25 degrees Centigrade. it can be performed under anhydrous conditions in the presence or absence of a catalyst, e. g., boron trifluoride; or under aqueous conditions in the presence or absence of a catalyst such as a hypohalous acid.

it anhydrous conditions are difiicult or inconvenient to maintain, the oxide opening reaction can be performed under aqueous conditions in which case the ketal will he hydrolyzed at the same time. The thus-produced 3-kelogroup of Ila can then be rckelalized in the manner described hereinabove to produce the corresponding kettil (III).

In the reduction step of the present invention, a 3- lLetalized 3 keto j-hydroxy-fidluoro-l l-oxygenatcib l7(20)-pregnene-2l-czirbonyloxy steroid (till, preferably the 3-ethyiene glycol kcial of loweralkyl, preferably methyl 3,l l-diketo-S-hyiiroxy-6-lluoro-l 7(2ll)-icisl-prc rnen-Zl-oute, is reduced with lithium aluminum hydride or other chemical carboxyl reducing agent in an organic solvent, 6. g, ether, dioxztric. tetrahydrofuran, benzene. to pro u the corresponding S-ketalized 5,] trill drilwdroxy- 6-Huoro-17(20)-pregnen-3-one. At completion of this reaction, the reaction mixture is preferably mixed with water or, an acid, an ester or carbonyl agent followed by water, to decompose any excess lithium aluminum hydride and organo-metal complexes. The usual reaction conditions for a lithium aluminum hydride reduction are employed, except that a reaction temperature at room temperature or below is preferred, to ensure that reaction with the G-fiuoro does not occur, and acid, though operative and satisfactory under carefully controlled conditions, is preferably not employed in the decomposition step, to avoid undue hydrolysis of the ketal group.

The esterification step, i. e., to produce compound V. involves the conversion of a ZI-hydroxy group of 5.11 5. 21-trihydroxy-6-fiuoro-l7(20)-pregnen-3-one B-alkylene ketal (IV) to a 2l-acyloxy group. This reaction can be performed under the esterification conditions known in the art, e. g., by the reaction of IV with the selected acid halide or acid chloride or acid bromide or the anhydride of a hydrocarbon carboxylic acid, or by reaction with the selected acid, in the presence of an esterification catalyst or with an ester under ester exchange reaction conditions. Reaction conditions which are apt to affect the labile llfi-hydroxy group or 6-fluoro group should be avoided. Compounds thus produced include the compounds represented by Formula V wherein the 17(20)- configuration is cis, and Ac is the acyl radical of a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive, e. g., formic, propionic, butyric, isobutyric, valeric, isovaleric, trimethylacetic, Z-methylbutyric, 3-ethylbutyric, hcxanoic, diethylacctic, triethylacetic, heptanoic, octanoic, a-ethylisovaleric, a cyclic acid, e. g., cyclopropylideneacetic, cyclopentylformic, cyclopentylacetic, B-cyclohexylpr-opionic, cyclohexylformic, cyclohexylacetic, an aryl or alkaryl acid, e. g., benzoic. 2-, 3- or 4-methylbenzoic, 2,3, 2,4-, 2,5-, 2,6-, 3,4-, and 3,5-dimethylbenzoic, ethylbenzoic, 2,4,6-trimethylbenzoic, 2,4,6-triethylbenzoic, a-naphthoic, 3-methyl-a-naphthoic, an aralkyl acid, e. g., phenylacetic, phenylpropionic, diphenylacetic, triphenylacetic, an unsaturated acid, e. g., acrylic, vinyl acetic, propiolic, undecolic, etc.

The oxidative hydroxylation of V to VI is carried out by reaction with a catalytic amount of osmium tetroxide and an oxidizing agent such as hydrogen peroxide, peracids, alkyl peroxides, amine oxide peroxides, and the like. A preferred procedure involves the use of two to three molar equivalents of one of the oxidizing agents and less than 0.05 molar equivalent of osmium tetroxide, calculated on the basis of V, using tertiary butyl alcohol as the reaction medium and carrying out the reaction at about room temperature.

The hydrolysis step involves the hydrolysis of the ketal group of the S-ketalized 5,1118,17a-trihydroxy-6-fiuoro- 2l-acyloxypregnane-3,20-dione, e. g., the ethylene glycol ketal thereof, employing aqueous acid to produce the corresponding diketo compound VII. The hydrolysis is conveniently performed under relatively mild conditions, e. g., at room temperature with acetic acid or dilute sulfuric acid.

The dehydration reaction is carried out using a mineral acid such as, for example, hydrogen chloride; or Girards reagent T. Acetic anhydride, p-toluenesulfonic acid and acetic acid, or thionyl chloride and pyridine are also suitable as dehydrating agents. Care should be taken in this step to avoid various reaction conditions that would affect the llfl-hydroxy group.

Compound VIII in the form of the 2l-ester can be oxidized to the corresponding ll-keto compound. i. e., 17a-hydroxy-6-fiuoro-2l-acyloxy 3,11,20 trione (compound IX), in accordance with the process for the oxidation of the 2l-csters of Kendalls compound F as disclosed in U. S. Patent No. 2,751,402.

Either of compounds VIII or IX can be transformed to the free 21-alcohol by hydrolysis with a base. A preferred procedure is to employ at least a molar equivalent of an alkali-metal bicarbonate or an alkali-metal carbonate in a substantially oxygen-free solution of a mixture of a lower alkanol and water. The hydrolysis reaction is carried out at a temperature between ten and thirty degrees centigrade while protecting the mixture from atmospheric oxygen. After the hydrolysis is complete, the reaction mixture is neutralized with an acid, e. g., acetic acid, and the hydrolyzed product recovered from the reaction mixture by evaporation and crystallization, extraction with methylene chloride, or the like.

As previously mentioned, the order of the various steps outlined above can be varied considerablyi. Suitable variations will occur to those skilled in the art, and the necessary adjustments can be readily appreciated and carried out by skilled chemists. For example, compound IV can be hydrolyzed to remove the 3-ketal, and this step followed in turn by 2l-acylation, S-dehydration and oxidative hydroxylation under the above-described conditions. In addition, ketal removal and dehydration of compound VI with, for example, hydrochloric acid in chloroform, is directly productive of compound VIII without isolation of. the intermediate compound VII.

The foregoing compounds, VIII or IX, all characterized by the presence of a 6-fiuoro substituent, can exist in either the 611- or the 6,9'epimeric form. The foregoing process produces a mixture in which the 6fl-form predominates. The fiix-epimer can be separated from the products by chromatographic or fractional crystallization techniques known in the art. The compounds are useful, however, as mixtures produced directly from the synthesis steps described. It is occasionally desirable, nevertheless, to obtain the fia-epimer from the mixture by separation or by epimerization. Conversion of the 6B- epimer or mixtures predominating therein can be accomplished by treatment at temperatures of zero degrees centigrade or slightly below in an organic solvent, such as chloroform, methylene chloride, ether, and the like, and in the presence of a prototropic agent (a protondonating reagent) such as water, alcohols, organic acids, and the like, with a mineral acid, such as, for example, hydrochloric acid. The mixture should be maintained at temperatures below zero degrees centigrade, or at least below room temperature during the addition of the acid. The reaction mixture can then be washed with successive portions of dilute alkali and water, and then dried and evaporated under reduced pressure. The fia-fluoro product can be recovered from the crude reaction product and purified by recrystallization.

The following examples are illustrative of the process and products of the present invention but are not to be construed as limiting.

EXAMPLE 1 The 3-ethylene ketal of methyl 3,11-diket0-5a,6a-oxido- 17(20) -[ci.r]-pregnen21 -0ate To a solution of 5.0 grams of the 3-ethylene glycol ketal of methyl 3,11-diketo4,17(20)-[cis]-pregnadien-21- oate, prepared in the manner described in U. S. Patent 2,707,184, in milliliters of chloroform was added a chilled solution of 1.9 grams of perbenzoic acid dissolved in 31.5 milliliters of chloroform. The solution was maintained at about four degrees centigrade for 24 hours, and then at room temperature for 72 hours. The solution was then washed with a five percent aqueous solution of sodium bicarbonate and then with water. The chloroform layer was separated, dried and the solvent distilled to give a residue of 5.3 grams of solid. Crystallization of this solid from methanol gave 2.24 grams of product melting at to degrees centigrade and after two crystallizations from methanol, there was obtained pure B-ethylene ketal of methyl 3,1l-diketo-5a,6otoxido-17(20)-[cisl-pregnen-Zl-oate melting at 206 to 209 degrees centigrade having a [0:1 of plus 37 degrees (CIiCla) and having the analysis given below:

Analysis.-Calculated for C H O C, 69.20; H, 7.75. Found: C, 69.59; H, 7.81.

EXAMPLE 2 To a solution of 1.73 grams of 3-ethylene ketal of methyl 3,11-diketo-5a,6a-oxido-1.7(29)-[cis]-pregnen-2loate in sixteen milliliters of methylene chloride was added six milliliters of 48 percent hydrofluoric acid. The heterogeneous mixture was stirred for two hours, made slightly basic with 300 milliliters of five percent sodium bicarbonate solution, and extracted with methylene chloride. The extract was washed, dried and evaporated to dryness to give 1.62 grams of crude solid. Chromatography gave two fractions: (A) 481 milligrams eluted with methylene chloride plus five percent acetone, and (B) 921 milligrams eluted with methylene chloride plus ten and twenty percent acetone. Crystallization of fraction A from acetone-Skellysolve B hexanes gave 390 milligrams of methyl 3,11-diketo-5a-hydroxy-6/3fluoro-17- (20)-allopregnen-21-oate, melting point 254 to 260 degress Centigrade. An analytical sample melted at 260 to 263 degrees Centigrade.

Analysis.Calculated for (1 11 1 F, 4.84. Found: F, 4.47.

Fraction B, on crystallization from acetone-Skellysolve B hexanes, gave 470 milligrams of methyl 3,11-diketo-5m; 6 3-dihydroxy-l7(20)-allopregnen-21-oate, melting point 235 to 245 degrees centigrade. An analytical sample melted at 245 to 248 degrees centigrade.

Analysis.-Calculated for C H O C, 67.67; H, 7.74. Found: C, 67.91; H, 7.62.

EXAMPLE 3 Methyl .5,II-diketo-5cx-hydroxyfidfluorr17(20) allopregnen-ZI-oate 3-ethylene ketal A mixture of 1.9 grams of methyl 3,11-diketo-5u-hydroxy 6B fluoro 17(20) allopregnen 21 oate, 59 milligrams of p-toluenesulfonic acid monohydrate and 31 milliliters of distilled ethylene glycol was added to 800 milliliters of benzene. The mixture was stirred and refiuxed for two hours, with the condensate passing through a water trap to remove the water. After reflux the mixture was cooled, washed with water and evaporated to dryness to give a crude solid which on recrystallization from acetone-Skellysolve B hexanes gave 1.96 grams of methyl 3,11 diketo 5a hydroxy 6e fluoro 17(20)- allopregnen-Zl-oate 3-ethylenc ketal, melting point 170 to 173 degrees Centigrade.

Following the above procedure, substituting other dihydric alcohols for ethylene glycol, for example, 1.2- propylene glycol, 2,3-butanediol, 1,3-butanediol and 2,3- pentanediol is productive of the respective 3-alkylene ketals of methyl 3,1l-diketo Swhydroxy-6fi-fluoro-l7(20)- allopregnen-Zl-oate.

EXAMPLE 4 To a solution of 1.96 grams of methyl 3,11-diketo-5ahydroxy 6B fiuoro 17(20) allopregnen 21 oate 3-ethylene ketal in 850 milliliters of anhydrous ether was added 3.7 grams of lithium aluminum hydride and the mixture was stirred for a period of one hour. 200 milliliters of Water was added slowly and the ether phase separated. The aqueous phase was extracted with ethyl acetate and the extracts added to the ether phase. The combined ether-cthyl acetate solution was Washed with water, dried and evaporated to dryness under reduced pressure. The crude solid residue was crystallized from acctoneSkellysolve B hexanes to give 1.30 grams of 511,115,21 trihydroxy 6; fluoro 17(20) allopregnen-3-one 3-ethylene ketal, melting point 197 to 205 degrees centigrade. An additional 226 milligrams was obtained from the mother liquor, melting point 175 to 185 degrees centigrade.

EXAMPLE 5 The acetate was prepared by allowing 0.87 gram of 501,] 15,21 trihydroxy 65 fluoro 17(20) allopregnen- 3one 3-ethylene ketal to stand overnight in ten milliliters of acetic anhydride and ten milliliters of pyridine. The solution was then poured into ice water to give 0.92 gram of 50 11 3 dihydroxy 6B fiuoro 21 acetoxy- 17(20) allopregnen 3 one 3 ethylene ketal, melting point to degrees centigrade, which on recrystaliization from acetone-Skellysolve B hexanes gave 0.77 gram, melting point 149 to 153 degrees centigrade.

Similarly, other 21-organic carboxylic esters of 5a,11e,21 trihydroxy 66 fiuoro 17(20) allopregnen-3-one 3-ethylene ketals are prepared wherein the 21- acyloxy group is formyloxy, propionyloxy, butyryloxy, valeryloxy, hexanoxyloxy, heptanoyloxy, octanoyloxy, benzyloxy, phenylacetoxy, or the like, by contacting 501,1 113,21 trihydroxy 6B fiuoro 17(20) allopregnen- 3-or1e 3-ethylene ketal with an appropriate acylating agent, e. g., the anhydride or acid halide of the selected acid in a solvent such as, for example, benzene, toluene, pyridine, or the like.

EXAMPLE 6 5051113,] 7m-trihydroxy-6e-flu0r0-21-acet0xyallopregnane- 3,20-drone S-ethylene ketal To a solution of 0.77 gram of 511,1149 dihydroxy 6/3- fluoro 21 acetoxy 17(20) allopregnen 3 one 3- ethylene ketal in 35 milliliters of tertiary butyl alcohol was added one milliliter of pyridine, 1.9 milliliters of N- methymorpholine oxide peroxide solution, and 13.1 milligrams of osmium tetroxide (9.1 milliliters of tertiary butyl alcohol solution containing 1.44 milligrams 0sO per milliliter). The solution was stirred for a period of 2.5 hours, fifteen milliliters of five percent sodium hydrosulfite added, stirred for an additional ten minutes, 0.7 gram of finely ground synthetic magnesium silicate added, stirred for a period of twenty minutes more and filtered. The filtrate was taken to dryness under reduced pressure (below fifty degrees Centigrade) and the residue dissolved in methylene chloride, washed with water, dried and evaporated to dryness. This residue was crystallized from acetoneSkellysolve B hexanes to give 0.47 gram of 5a,11p3,17u trihydroxy 6B fluoro 21 acetoxyallopregnane 3,20 dione 3 ethylene ketal, melting point 220 to 228 degrees Centigrade.

EXAMPLE 7 A solution of 0.47 gram of 5a,llB,17a trihydroxy- 6p fluoro 21 acetoxyallopregnane 3,20 dione 3- ethylene ketal in 35 milliliters of acetone and four milliliters of 1 N sulfuric acid solution was gently boiled on the steam bath for ten minutes, cooled and neutralized with dilute sodium bicarbonate solution. Addition of water and cooling gave 0.33 gram of 5u,11;S,l7a-trihydroxy 6,1-1- flucro 21 acetoxyallopregnane 3,20 dionc, melting point 230 to 240 degrees centigrade.

A solution of 100 milligrams of 5 s.] lfl,l7r--trihydroxy 6/3 iluoro 21 acetoxyallopregnane 3,20 dione in 4.9 milliliters of acetic acid and 0.1 milliliter of water was refluxed for a period of cue hour, cooled, diluted with fifty milliliters of water and evaporated to dryness under reduced pressure. The residue was chromatographed over Florisil (synthetic magnesium silicate) to give one fraction (77 milligrams) eluted with methylene chloride plus ten percent acetone. Crystallization from acetone- Skellysolve B hexanes gave 38 milligrams of 6B fluorol1fl,17on dihydroxy 21 acetoxy 4 pregnene 3,20- dione (6,6-fluorohydrocortisone acetate), melting point 210 to 218 degrees centigrade. Infrared data. and ultraviolet data were found to be in agreement with the structure.

EXAMPLE 9 Isomerization of 618- to 6m-flu0rohydr0c0rtis0ne acetate A solution of 0.132 gram of GB-fluorohydrocortisone acetate in twelve milliliters of chloroform and 0.1 milliliter of absolute alcohol was cooled to minus ten degrees Centigrade in an ice-salt bath and .a stream of anhydrous hydrochloric acid was gently bubbled through the solution for 2.5 hours while the temperature was maintained between minus five and minus fifteen degrees centigrade. The solution was then diluted with 25 milliliters of chloroform, washed with dilute sodium bicarbonate and water, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure at sixty degrees centigrade or less. Crystallization of the residue from acetone- Skellysolve B gave 42 milligrams of product, 6a-fluorohydrocortisone acetate, melting point 203 to 210 degrees centigrade.

EXAMPLE 10 hydroxy-ZI-acetoxy-4-pregnene-3,1I ,20-trine (6m-flu0- rocortisone acetate) Oxidation of 6(3-flu0rohydrocortisone acetate and 6afluorohydrocortisone acetate of Examples 8 and 9 with 5 A solution of 1.1 grams of 6a-fluorohydrocortisone acetate, 1.0 gram of potassium bicarbonate, 100 milliliters of methanol and fifteen milliliters of water was purged with nitrogen and stirred at degrees centigrade for four hours. The solution was then neutralized by addition of acetic acid and the methanol was removed by distillation under reduced pressure. The residue was extracted with 100 milliliters of methylene dichloride and the extract, after drying over sodium sulfate, was chromatographed over a column of eighty grams of synthetic magnesium silicate. The product fraction was eluted with Skellysolve B hexanes plus twenty and thirty percent acetone and gave 770 milligrams of 6a-fluorohydrocortisone which melted at 192 to 195 degrees centigrade after crystallization from ethyl acetate-Skellysolve B hexanes. An analytical sample melted at 192 to 201 degrees centigrade and had a rotation of [al plus 127 degrees (chloroform).

Analysis.-Calcd. for C H O F: C, 66.29; H, 7.68; F, 4.99. Found: C, 66.28; H, 7.65; F, 4.43.

Following the procedure of Example 11, above, saponification of 6fl-fluorohydrocortisone acetate, 6fl-fluorocortisone acetate and 6u-fluorocortisone acetate of Examples 8 and 10 is productive of the corresponding 21-hydroxy compounds, 6,9-fluorohydrocortisone, 6B-fluorocortisone and 6a-iluorocortisone.

As previously described, the 21-acyloxy products of this invention, e. g., the products of Examples 8, 9 and 10 can be hydrolyzed to the corresponding free 21-alc0hols by the hydrolysis procedure described above. In the hydrolysis procedure, as previously stated, it is desirable to employ an oxygen-free solution and oxygen-free conditions and to use a molar excess of an alkali-metal bicarbonate such as potassium bicarbonate as a hydrolytic 10 agent. The temperature is preferably held lower, i. e., between ten and about thirty degrees centigrade, and the hydrolyzed solution subsequently neutralized with an acid such as acetic acid.

EXAMPLE 12 Five -milliliter portions of a medium, in 250-milliliter Erlenmeyer flasks, containing one percent glucose, two percent corn steep liquor (sixty percent solids) and tap water, were adjusted to a pH of 4.9. This medium was sterilized for 45 minutes at fifteen pounds per square inch pressure and inoculated with a one to two day growth of Septomyxa afiinis, A. T. C. C. 6737. The Erlenmeyer flask was shaken at room temperature (about 26 to 28 degrees centigrade) for a period of three days. At the end of this period this SOD-milliliter volume was used as an inoculum for ten liters of the same glucose-corn steep liquor medium which in addition contained ten milliliters of an antifoam compound (a mixture of lard oil and octadecanol). The fermenter was placed into the waterbath, adjusted to 28 degrees centigrade and the contents stirred (300 R. P. M.) and aerated (0.3 liter air per minute to five liters of beer). After 24 hours of incubation, when a good growth had been developed, five grams of Gu-flUOI'OhYdl'OCOI'ilSOIlC acetate plus one-half gram of 3-ketobisnor-4-cholen-22-al, dissolved in 25 milliliters of dimethylformamide was added and the incubation carried out at the same temperature (28 degrees centigrade) and aeration for a period of 72 hours (final pH 8.3). The mycelium was filtered off and washed with water. The wash water was combined with the filtrate and the whole was extracted with three two-liter portions of a mixture of methylene-ethyl acetate (3:1). Removal of the solvent by evaporation gave 5.25 grams of crude solid which was triturated twice with four milliliters of methylene chloride to give 2.4 grams of 1-dehydro-6a-fiuorohydro cortisone of melting point 198 to 203 degrees centigrade. The analytical sample, recrystallized from acetone, melted at 202 to 204 degrees centigrade. Analysis gave [(11 plus 73 degrees (dioxane) and the following:

Analysis.Calcd. for C H O F: C, 66.65; H, 7.10; F, 5.02. Found: C, 66.68; H, 7.19; F, 5.49.

Similarly, fermentation of 6a-fluorocortisone acetate with Septomyxa afiinis is productive of l-dehydro-6afluorocortisone It is to be understood that the invention is not to be limited to the exact details of operation or exact compounds shown and described, as obvious modifications and equivalents will be apparent to one skilled in the art, and the invention is therefore to be limited only by the scope of the appended claims.

We claim:

1. 6-fluoro-1l-oxygenated-lh, 2l-dihydroxy- 4 pregnene-3,20-dione of the following formula:

omoH

wherein R is selected from the group consisting of hydroxy and keto, and the 21-acy1ates thereof wherein the acyl radical is that of a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive.

2. 6-fluorohydrocortisone and the 21-acylates thereof wherein the acyl radical is that of a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive.

vapo -lacuna.

wherein R is selected from the group consisting of hydroxy and keto, R and R" are selected from the group consisting of hydrogen and lower-alkyl, and n is a whole number from one to two.

14. The 3-ethylene ketal of methyl 3,l.l-dikt-5a,6otoxido-l7(2t))-[cis] -pregnen-21-oate.

15. A compound selected from the group consisting of 3 keto oz hydroxy 6B fiuoro-l1-0xygenated-17(2D)- pregnene-Zl-carbonyloxy steroid represented by the forrnula:

O and the 3-cyclic ketals thereof represented by the follow ing formula:

no 0 R on i R CH: 1

l on o,

y (cuntwherein R is selected from the group consisting of hy' droxy and lzeto, R and R" are selected from the group consisting of hydrogen and lower-alkyl, and n is a whole number from one to two.

Fili) 16. The 3-ethylene keta] of methyl 3,1l-diketo-5a-hydroXy-6B-fluoro-17(20)-allopregnen-21-oate.

17. Methyl 3,11 diketo-Sa-hydroXy-Gfl-fluOro-17(20)- allopregnen-2l-oate.

l8. S-ketalized 3 -keto-5a,llB,2l-trihydroxy-6fi-fluorol7(20)-allopregnene represented by the following formula:

wherein R" is selected from the group consisting of hydrogen and lower alkyl, 11 is a whole number from one to two, and Zl-acylates thereof wherein the acyl radical is that of a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive.

19. The 3-ethylene ketal of 5a,ll,8,2l-trihydroxy-6ti fiuoro-l 7 (20) allopregnen-3one.

20. The 3ethylene ketal of 5o 1lfi-dihydroxy-Zl-acetoxy-6/8-fiuorol7 20) -allopregnen-3-one.

21. A compound selected from the group consisting of 511,1 1 a, l 7a-trihydroxy-21-acyloxy-6fi-fluoroallopregnan-3- one represented by the following formula:

onZ

onion 0:0

and 3-cyclic ketals thereof represented by the following formula:

tP U J tem wherein R" is selected from the group consisting of hydrogen and lower alkyl, n is a whole number from one to two, and Ac is the acyl radical of a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive.

22. The B-ethylene ketal of 511,11fi,17a-trihydroxy-21- acetoxy-6B-fluoroallopreg11an-3,ZO-dione.

23. 5 x,l 15,170. trihydroxy 2i acetoxy-fifi fluoroallopregnan-3,20--dione.

24. A process for the production of d-flnoro-IIBJM- dihydroxy-Zl-acyloxy 4-pregnene-3,20-dione of the following formula:

wherein Ac is the acyl radical of an organic carboxylic acid containing from one to twelve carbon atoms, inclusive, which comprises: (1) epoxidizing a compound of the following formula:

wherein R is selected from the group consisting of hydroxy and keto, R and R" are selected from the group consisting of hydrogen and lower alkyl, and n is a whole number from one to two, with an epoxidizing agent to form the corresponding Super-oxide; (2) reacting the said a,6a-oxide with a fluorinating agent to produce the corresponding 3-keto-5a-hydroxy-6B-fiuoro-ll-oxygenated- 17(20)-allopregnene-2l-carbonyloxy compound; (3) kctalizing the said 3-keto-6-fiuoro compound to produce the corresponding 3-ketal; (4) reducing the said 3-ketalized compound with a carboxyl reducing agent in an organic reaction medium to produce the corresponding 3-ketalized 5 a,l 1fi,2 1 -trihydroxy-6[3-fluorol 7 (20) -a1lopregnen-3-one; (5) esterifying the said 3-ketalized 11,8,21- dihydroxy compound with an esterifying agent to produce the corresponding 2l-ester; (6) oxidatively hydroxylating the said 21-ester with osmium tetroxide and an oxygen-donating oxidizing agent to produce the corresponding B-ketalized 17:1-hYdI0XY compound; (7) deketalizing the said 3-ketalized 17a-hydroxy compound by hydrolysis to produce the corresponding 3-keto compound, and (8) dehydrating the said 3-keto compound to produce 6-fluoro-11,6,l7a-dihydroxy-21-acyloxy-4- pregnene-3 ,ZO-dione.

25. A process for the production of compounds of the following formula:

C 0 O R R! Eli-o ahi I '0 wherein R is a member selected from the group consisting of hydroxy and keto, R and R" are members selected from the group consisting of hydrogen and lower alkyl,

and n is a whole number from one to two, which comprises: epoxidizing a compound of the following formula:

C O O R wherein R, R, R" and n are defined as above, with an epoxidizing agent to produce the said 5,6-oxido compound.

26. The process of claim 25 wherein the epoxidizing agent is a member selected from the group consisting of perbenzoic acid and peracetic acid.

27. A process for the production of compounds of the following formula:

RI! orb-o (bum-o 5 wherein R is a member selected from the group consisting of hydroxy and keto, R and R" are members selected from the group consisting of hydrogen and lower alkyl, n is a whole number from one to two, which comprises: reacting a compound of the following formula:

wherein R, R, R and n are defined as above, with a fluorinating agent to produce the corresponding 3-keto- 5a-hydroxy-6fi-fluoro ll oxygenated-17(20)-pregnene- 21-carbonyloxy compound, and reacting the said 3-keto compound with a ketalizing agent.

28. The process of claim 27 wherein the fluorinating agent is hydrofluoric acid and the ketalizing agent is ethylene glycol in the presence of an acid catalyst.

29. A process for the production of compounds of the following formula:

CHI

CHsOAc OH; R! iH-O H ..-o i 2) OH (IJOOR OH l CH-- I (cum-0 OHl wherein R is a member selected from the group consisting of hydroxy and keto, R and R are members selected from the group consisting of hydrogen and lower alkyl, and It is defined as above, with a reducing agent to produce the corresponding 3-ketalized 3-kcto-Sa,1l}8,21-trihydroxy-6rl-fiuoro-l7(20)-allopregncne, and esterifying the said trihydroxy compound with an esterifying agent.

30. The process of claim 29 wherein the reducing agent is lithium aluminum hydride and the esterifying agent is a member selected from the group consisting of acid anhydrides and acid halides of organic carboxylic acids containing from one to twelve carbon atoms, inelusive.

31. A process for the production of compounds of the following formula:

01110 R 0:0 |---OH wherein R' is a member selected from the group consisting of hydrogen and the acyl radical of an organic carboxylic acid containing from one to twelve carbon atoms, inclusive, which comprises: oxidatively hydroxylating a 3 ketalized 3-kcto-5a,llB-dihydroxy-Zl-acyloxy- 65-fiuoro-l7(20)-allopregnene, with osmium tetroxide and an oxidizing agent, wherein the acyl radical is defined as above, to produce the corresponding 3-ketalized 17ahydroxy compound; deketalizing the said 3-ketalized 17ahydroxy compound by hydrolysis to produce the corresponding 3-keto compound, and dehydrating the said 3-keto compound with a mineral acid.

32. The process of claim 31 wherein the oxidative hydroxylating agent is osmium tetroxide and an amine oxide peroxide, hydrolysis is by an acid hydrolyzing agent, and the dehydrating agent is acetic acid.

33. A process for the production of compounds of the following formula:

CHqOAe :0 on

wherein Ac is the acyl radical of an organic carboxylic acid containing from one to twelve carbon atoms, inclusive, which comprises: reacting a compound of the formula:

wherein Ac is as above defined, R" is selected from the group consisting of hydrogen and lower-alkyl, and n is a whole number from one to two, with a mineral acid.

34. A process for the production of compounds of the following formula:

crnoac ona wherein Ac is the acyl radical of an organic carboxylic acid containing from one to twelve carbon atoms, inclusive, which comprises: reacting a compound of the formula:

wherein Ac is defined as above, R" is selected from the group consisting of hydrogen and lower-alkyl, and n is a whole number from one to two, with hydrochloric acid.

35. A process for the isomerization of a GB-fluoro compound of the following formula:

onto a 0:0 ---orr wherein R is selected from the group consisting of hydro gen and the acyl radical of an organic carboxylic acid containing from one to twelve carbon atoms, inclusive, to the corresponding 6a-fluoro compound, which comprises: reacting the 6B-fluoro epimer with a mineral acid in the presence of a prototropic agent to obtain the wherein R is selected from the group consisting of hycorresponding Ga-fiuoro epimer. drogen and the acyl radical of an organic carboxylic acid 36. A process for the isomerization of the GB-fluoro containing from one to twelve carbon atoms, inclusive, compound of the following formula: to the corresponding 6u-fiuoro compound, which com- 5 prises: reacting the 6l8-fluoro epimer with hydrochloric (HJHZOR acid in the presence of an alcohol to obtain the corresponding a-fiuoro epimer. HO References Cited in the file of this patent OH 10 UNITED STATES PATENTS 3 2,707,184 Hogg et al. Apr. 26, 1955 2,742,461 Bernstein et a1 Apr. 17, 1956 2,781,368 Heyl et al. Feb. 12, 1957 0 15 OTHER REFERENCES Mattox et al.: J. Biol. Chem. 197 (1952), page 263.

UNITED STATES PATENT OFFICE Certificate of Correction Patent No. 2,838,497 June 10, 1958 George B. Spero, at 1:1.

It is horeby certified that error appears in the printed specification of the above numbered about requiring correction and that the said Letters Patent should read as correote below.

Column 7, line 7, for -17(29)- read -17 (20} column 8, line 37, for methymorpholine read -methy]morpho1ino-; column 12, line 34, for 11 read 11,6-.

Signed and sealed this 17th day of March 1959.

Atfash KARL H. AXLINE, ROBERT C. WATSON,

Amati/21g Oflicer. Commissioner of Patents. 

1. 6-FLUORO-11-OXYGENATED-17A,21-DIHYDROXY-4-PREGNENE-3,20-DIONE OF THE FOLLOWING FORMULA:
 13. A 3-KETALIZED 3-KETO-5A,6A-OXIDO-11-OXYGENATED-17(20)-PREGNENE-21-CARBONYLOXY STERIOD REPRESENTED BY THE FOLLOWING FORMULA: 